tetano
Editor, Senior Moderator
Int J Mol Sci
. 2022 Apr 6;23(7):4063.
doi: 10.3390/ijms23074063.
In Vitro SARS-CoV-2 Infection of Microvascular Endothelial Cells: Effect on Pro-Inflammatory Cytokine and Chemokine Release
Maria Dolci[SUP] 1 [/SUP], Lucia Signorini[SUP] 1 [/SUP], Sarah D'Alessandro[SUP] 2 [/SUP], Federica Perego[SUP] 1 [/SUP], Silvia Parapini[SUP] 3 [/SUP], Michele Sommariva[SUP] 4 [/SUP], Donatella Taramelli[SUP] 2 [/SUP], Pasquale Ferrante[SUP] 1 [/SUP], Nicoletta Basilico[SUP] 1 [/SUP], Serena Delbue[SUP] 1 [/SUP]
Affiliations
Abstract
In the novel pandemic of Coronavirus Disease 2019, high levels of pro-inflammatory cytokines lead to endothelial activation and dysfunction, promoting a pro-coagulative state, thrombotic events, and microvasculature injuries. The aim of the present work was to investigate the effect of SARS-CoV-2 on pro-inflammatory cytokines, tissue factor, and chemokine release, with Human Microvascular Endothelial Cells (HMEC-1). ACE2 receptor expression was evaluated by western blot analysis. SARS-CoV-2 infection was assessed by one-step RT-PCR until 7 days post-infection (p.i.), and by Transmission Electron Microscopy (TEM). IL-6, TNF-α, IL-8, IFN-α, and hTF mRNA expression levels were detected by RT-PCR, while cytokine release was evaluated by ELISA. HMEC-1 expressed ACE2 receptor and SARS-CoV-2 infection showed a constant viral load. TEM analysis showed virions localized in the cytoplasm. Expression of IL-6 at 24 h and IFN-α mRNA at 24 h and 48 h p.i. was higher in infected than uninfected HMEC-1 (p < 0.05). IL-6 levels were significantly higher in supernatants from infected HMEC-1 (p < 0.001) at 24 h, 48 h, and 72 h p.i., while IL-8 levels were significantly lower at 24 h p.i. (p < 0.001). These data indicate that in vitro microvascular endothelial cells are susceptible to SARS-CoV-2 infection but slightly contribute to viral amplification. However, SARS-CoV-2 infection might trigger the increase of pro-inflammatory mediators.
Keywords: Human Microvascular Endothelial Cells (HMEC-1); SARS-CoV-2; inflammatory mediators.
. 2022 Apr 6;23(7):4063.
doi: 10.3390/ijms23074063.
In Vitro SARS-CoV-2 Infection of Microvascular Endothelial Cells: Effect on Pro-Inflammatory Cytokine and Chemokine Release
Maria Dolci[SUP] 1 [/SUP], Lucia Signorini[SUP] 1 [/SUP], Sarah D'Alessandro[SUP] 2 [/SUP], Federica Perego[SUP] 1 [/SUP], Silvia Parapini[SUP] 3 [/SUP], Michele Sommariva[SUP] 4 [/SUP], Donatella Taramelli[SUP] 2 [/SUP], Pasquale Ferrante[SUP] 1 [/SUP], Nicoletta Basilico[SUP] 1 [/SUP], Serena Delbue[SUP] 1 [/SUP]
Affiliations
- PMID: 35409421
- DOI: 10.3390/ijms23074063
Abstract
In the novel pandemic of Coronavirus Disease 2019, high levels of pro-inflammatory cytokines lead to endothelial activation and dysfunction, promoting a pro-coagulative state, thrombotic events, and microvasculature injuries. The aim of the present work was to investigate the effect of SARS-CoV-2 on pro-inflammatory cytokines, tissue factor, and chemokine release, with Human Microvascular Endothelial Cells (HMEC-1). ACE2 receptor expression was evaluated by western blot analysis. SARS-CoV-2 infection was assessed by one-step RT-PCR until 7 days post-infection (p.i.), and by Transmission Electron Microscopy (TEM). IL-6, TNF-α, IL-8, IFN-α, and hTF mRNA expression levels were detected by RT-PCR, while cytokine release was evaluated by ELISA. HMEC-1 expressed ACE2 receptor and SARS-CoV-2 infection showed a constant viral load. TEM analysis showed virions localized in the cytoplasm. Expression of IL-6 at 24 h and IFN-α mRNA at 24 h and 48 h p.i. was higher in infected than uninfected HMEC-1 (p < 0.05). IL-6 levels were significantly higher in supernatants from infected HMEC-1 (p < 0.001) at 24 h, 48 h, and 72 h p.i., while IL-8 levels were significantly lower at 24 h p.i. (p < 0.001). These data indicate that in vitro microvascular endothelial cells are susceptible to SARS-CoV-2 infection but slightly contribute to viral amplification. However, SARS-CoV-2 infection might trigger the increase of pro-inflammatory mediators.
Keywords: Human Microvascular Endothelial Cells (HMEC-1); SARS-CoV-2; inflammatory mediators.